Structural and Mechanical Hierarchies in the α-Crystallin Domain Dimer of the Hyperthermophilic Small Heat Shock Protein Hsp16.5

Morten Bertz, Jin Chen, Matthias J. Feige, Titus M. Franzmann, Johannes Buchner, Matthias Rief

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

In biological systems, proteins rarely act as isolated monomers. Association to dimers or higher oligomers is a commonly observed phenomenon. As an example, small heat shock proteins form spherical homo-oligomers of mostly 24 subunits, with the dimeric α-crystallin domain as the basic structural unit. The structural hierarchy of this complex is key to its function as a molecular chaperone. In this article, we analyze the folding and association of the basic building block, the α-crystallin domain dimer, from the hyperthermophilic archaeon Methanocaldococcus jannaschii Hsp16.5 in detail. Equilibrium denaturation experiments reveal that the α-crystallin domain dimer is highly stable against chemical denaturation. In these experiments, protein dissociation and unfolding appear to follow an "all-or-none" mechanism with no intermediate monomeric species populated. When the mechanical stability was determined by single-molecule force spectroscopy, we found that the α-crystallin domain dimer resists high forces when pulled at its termini. In contrast to bulk denaturation, stable monomeric unfolding intermediates could be directly observed in the mechanical unfolding traces after the α-crystallin domain dimer had been dissociated by force. Our results imply that for this hyperthermophilic member of the small heat shock protein family, assembly of the spherical 24mer starts from folded monomers, which readily associate to the dimeric structure required for assembly of the higher oligomer.

Original languageEnglish
Pages (from-to)1046-1056
Number of pages11
JournalJournal of Molecular Biology
Volume400
Issue number5
DOIs
Publication statusPublished - 2010 Jul
Externally publishedYes

Fingerprint

Small Heat-Shock Proteins
Crystallins
Methanocaldococcus
Protein Unfolding
Molecular Chaperones
Archaea
Proteins

Keywords

  • α-crystallin domain
  • Dissociation force
  • Force spectroscopy
  • Protein stability
  • Small heat shock proteins

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Structural and Mechanical Hierarchies in the α-Crystallin Domain Dimer of the Hyperthermophilic Small Heat Shock Protein Hsp16.5. / Bertz, Morten; Chen, Jin; Feige, Matthias J.; Franzmann, Titus M.; Buchner, Johannes; Rief, Matthias.

In: Journal of Molecular Biology, Vol. 400, No. 5, 07.2010, p. 1046-1056.

Research output: Contribution to journalArticle

Bertz, Morten ; Chen, Jin ; Feige, Matthias J. ; Franzmann, Titus M. ; Buchner, Johannes ; Rief, Matthias. / Structural and Mechanical Hierarchies in the α-Crystallin Domain Dimer of the Hyperthermophilic Small Heat Shock Protein Hsp16.5. In: Journal of Molecular Biology. 2010 ; Vol. 400, No. 5. pp. 1046-1056.
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